Effects Of Nodal Gene RNA Interference On Biological Behaviour And Vasculogenic Mimicry Of Human Hepatocellular Carcinoma Cells

Backgroud Hepatocellular carcinoma is one of the most common malignancy in theworld, and is a major challenge for the clinical treatment especially in the high incidencerate of postoperative recurrence and metastasis. Hepatocellular carcinoma is the result ofthe interaction of multiple genes, multiple stages and multiple factors. The differentstages of oncogenes or anti-oncogenes variation in the genesic and development processcause related factors expression disorder. Nodal is a TGF-β supper family member.Existing relevant studies have shown that there are closely associativity between Nodalexpression and tumor. Thus, the study of relationship between Nodal and hepatocellularcarcinoma has great significance to understand the genesis and development ofhepatocellular carcinoma and find effective treatment methods.Objective To silence the expression of nodal gene in human hepatocellular carcinomacells by RNA interference (RNAi), and to observe the effect on biological behaviourand vasculogenic mimicry of them. Then, to explore the availability of nodal gene inpredicting metastasis and recurrence and targeted therapy in hepatocellular carcinoma. Methods4expression vectors of nodal gene targeting small interference RNA wereconstructed and transfected into human hepatocellular carcinoma cells SMMC-7721.Transfection efficiency was observed after the plasmid that with green fluorescentprotein gene had been transfected into target cells. Real-time quantitive PCR andWestern blot were used to examine nodal gene expression in all cancer cells. CCK-8assay was used to detect cell proliferation; Annexin V/PI apoptosis detection kit andflow cytometry were used to detect cell apoptosis; Transwell assay was used to detectcell capability of infestation and migration;3D cultivation was used to observevasculogenic mimicry of hepatocellular carcinoma cells. The Effects of nodal geneRNA interference on proliferation, apoptosis, infestation, migration and vasculogenicmimicry of human hepatocellular carcinoma cells were studed.Results There is a good transfection efficiency that Nodal gene specific siRNAplasmid transfected into human hepatocellular carcinoma cells SMMC-7721, whichcould reach about70%. The expression of nodal gene was suppressed effectively inhuman hepatocellular carcinoma cells by RNA interference. In the first4,5,6days ofprolieration experiment, the proliferation of interference group was sigenificantly lowerthan the control group(p <0.05);48h after transfection, the apoptosis rate ofinterference group was significantly higher than the control group(p <0.05); In theinfestation and migration experiments, the cells through the transwell chamber in theinterference group were lower than the control group(p <0.05);24h and48h after transfection, the vasculogenic mimicry in the interference group did not form,significantly different from the control group.Conclusions Nodal gene expression of human hepatocellular carcinoma cellsSMMC-7721had been successfully inhibited by RNA interference. Interferencing theexpression of nodal gene could inhibit the biological behaviour and the formation ofvasculogenic mimicry in human hepatocellular carcinoma cells. Therefore, it wasexpected to be a new ideal target of gene-specific therapy for hepatocellular carcinoma.